• 제목/요약/키워드: Assay system

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Biomonitoring the Genotoxicity of Environmental Pollutants Using the Tradescantia Bioassay (환경 중 유전독성물질 검색을 위한 자주달개비 생물검정 기법의 적용연구)

  • 신해식
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2004.05a
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    • pp.47-60
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    • 2004
  • Higher plants can be valuable genetic assay systems for monitoring environmental pollutants and evaluating their biological toxicity. Two assays are considered ideal for in situ monitoring and testing of soil, airborne and aqueous mutagenic agents; the Tradescantia stamen hair assay for somatic cell mutations and the Tradescantia micronucleus assay for chromosome aberrations. Both assays can be used for in vivo and in vitro testing of mutagens. Since higher plant systems are now recognized as excellent indicators and have unique advantages over in situ monitoring and screening, higher plant systems could be accepted by regulatory authorities as an alternative first-tier assay system for the detection of possible genetic damages resulting from the pollutants or chemicals used and produced by industrial sectors. It has been concluded that potential mutagen and carcinogen such as the heavy metals among indoor air particulates, volatile compounds in the working places, soil, and water pollutants contribute to the overall health risk. This contribution can be considerable under certain circumstances. It is therefore important to identify the level of genotoxic activity in the environment and to relate it to the biomarkers of a health risk in humans. The results from the higher plant bioassays could make a significant contribution to assessing the risks of pollutants and protecting the public from agents that can cause mutation and/or cancer. The plant bioassays, which are relatively inexpensive and easy to handle, are recommended for the scientists who are interested in monitoring pollutants and evaluating their environmental toxicity to living organisms.

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Assessment of Environmental Pollution with Tradescantia Bioassays (자주달개비 생물검정 기법을 이용한 환경오염 평가)

  • Kim Jin Gyu;Sin Hae Sik
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2004.05a
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    • pp.1-15
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    • 2004
  • Higher plants can be valuable genetic assay systems for monitoring environmental pollutants and evaluating their biological toxicity. Two assays are considered ideal for in situ monitoring and testing of soil, airborne and aqueous mutagenic agents; the Tradescantia stamen hair assay for somatic cell mutations and the Tradescantia micronucleus assay for chromosome aberrations. Both assays can be used for in vivo and in vitro testing of mutagens. Since higher plant systems are now recognized as excellent indicators and have unique advantages over in situ monitoring and screening, higher plant systems could be accepted by regulatory authorities as an alternative first-tier assay system for the detection of possible genetic damages resulting from the pollutants or chemicals used and produced by industrial sectors. It has been concluded that potential mutagen and carcinogen such as the heavy metals among indoor air particulates, volatile compounds in the working places, soil, and water pollutants contribute to the overall health risk. This contribution can be considerable under certain circumstances. It is therefore important to identify the level of genotoxic activity in the environment and to relate it to the biomarkers of a health risk in humans. The results from the higher plant bioassays could make a significant contribution to assessing the risks of pollutants and protecting the public firom agents that can cause mutation anuor cancer. The plant bioassays, which are relatively inexpensive and easy to handle, are recommended for the scientists who are interested in monitoring pollutants and evaluating their environmental toxicity to living organisms.

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Antioxidant and Antiproliferative Activities of Methanolic Extract from Celandine

  • Hu, Weicheng;Wang, Myeong-Hyeon
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.207-212
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    • 2009
  • Celandine (Chelidonium majus, family Papaveraceae) is an herb used extensively in traditional Korean medicine. To investigate its antioxidant and antiproliferative activities, the methanolic extract of celandine was introduced. The antioxidant properties of the extract were tested using various in vitro systems, including hydroxyl radical scavenging assay, DNA damage protection assay, 1,1-diphenyll-2-2-pricylhydrazyl (DPPH) free radical scavenging activity, metal chelating activity, and reducing power assay. The extract exhibited stronger antioxidant activity ($IC_{50}=7.92{\mu}g/mL$) against hydroxyl radicals in the Fenton system than butylated hydroxyanisole ($IC_{50}=51.46{\mu}g/mL$) and $\alpha$-tocopherol ($IC_{50}=67.48{\mu}g/mL$). Likewise, damage to the plasmid pBR 322 induced by hydroxyl radicals was found to be protected by the extract at a concentration of $400{\mu}g/mL$. Cellular proliferation and the induction of apoptosis were also examined by a cellular proliferation assay, flow cytometry, and mRNA expression analysis. Taken together, the extract significantly inhibited the growth of HT-29 cells in a concentration- and time-dependent manner, and gradually increased both the proportion of apoptotic cells and the expression of caspase-3. Overall, our research suggests that celandine possesses antioxidant and antiproliferative properties.

The antimutagenic effect and genetic safety of Buthus martensi Karsch aqua-acupuncture solution (BMKAS) (전갈(全蝎) 약침액(藥鍼液)의 항돌연변이(抗突然變異) 및 항암(抗癌) 효과(效果))

  • Kim, So-Houng;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.17 no.3
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    • pp.151-167
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    • 2000
  • Objective : The aim of this study is to determine the antimutagenic effect and genetic safety of Buthus martensi Karsch aqua-acupuncture solution(BMKAS) against various chemical carcinogens. Method : Ames(Salmonella typhimurium) test and Rec assay(Bacillus subtilis) were used as indicators for DNA damage and antimutagenesis. Furthermore, the levels of umu operon expression by measuring the ${\beta}$-galactosidase activity wete monitored with the SOS umu test using S. typhimurium 1535 containing plasmid pSK1002. And the host-mediated assay was used to investigate the mutagenicity and antimutagenicity of BMKAS inducing various chemical carcinogens after the activation with in vivo metabolic systems. Results : From the results, BMKAS did not atfect DNA of S. typhimurium and B. subtilis strains and showed no mutagenicity at the all concentrations of tested solution. Furthermore BMKAS dose-dependently protected the mutagenecity by AF-2, 2-AA and B[a]P. These phenomena was also similar to that after metabolic activation of BMKAS in in vivo system. Conclusion : These results suggested that BMKAS did not show the mutagenicity and protected the mutagenesis against various chemical carcinogens by four different methods used in this study.

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A MALDI-MS-based Glucan Hydrolase Assay Method for Whole-cell Biocatalysis

  • Ahn, Da-Hee;Park, Han-Gyu;Song, Won-Suk;Kim, Seong-Min;Jo, Sung-Hyun;Yang, Yung-Hun;Kim, Yun-Gon
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.69-77
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    • 2019
  • Screening microorganisms that can produce glucan hydrolases for industrial, environmental, and biomedical applications is important. Herein, we describe a novel approach to perform glucan hydrolase screening-based on analysis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) spectra-which involves degradation of the oligo- and polysaccharides. As a proof-of-concept study, glucan hydrolases that could break down glucans made of several glucose units were used to demonstrate the MALDI-MS-based enzyme assay. First, the enzyme activities of ${\alpha}$-amylase and cellulase on a mixture of glucan oligosaccharides were successfully discriminated, where changes of the MALDI-MS profiles directly reflected the glucan hydrolase activities. Next, we validated that this MALDI-MS-based enzyme assay could be applied to glucan polysaccharides (i.e., pullulan, lichenan, and schizophyllan). Finally, the bacterial glucan hydrolase activities were screened on 96-well plate-based platforms, using cell lysates or samples of secreted enzyme. Our results demonstrated that the MALDI-MS-based enzyme assay system would be useful for investigating bacterial glucoside hydrolases in a high-throughput manner.

Establishment and Application of a Multibead Serotyping Assay for Pneumococci in Korea (다중구슬 폐구균 혈청형 분석법의 국내 확립과 적용 연구)

  • Kim, Han Wool;Lee, Soyoung;Lee, Miae;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.22 no.2
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    • pp.97-105
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    • 2015
  • Purpose: Serotyping pneumococcal isolates is important to monitor efficacy of pneumococcal vaccines. Because of difficulties of typing pnueumocci, a multiplex bead-based (multibead) serotyping assay was recently introduced. The aim of this study is to establish a new multibead serotyping assay and to apply this method to analyze clinical isolates of pneumococci in Korea. Methods: To establish the multibead serotyping assay, six key reagents were transferred from University of Alabama at Birmingham (UAB) to Ewha Center for Vaccine Evaluation and Study (ECVES): bead set coated with polysaccharide and monoclonal antibody pool were used in one multiplex inhibition-type immunoassay and 2 bead sets coated DNA probe and 2 primer pools were used in two multiplex PCR-based assays. After multibead serotyping assay was set up, 75 test samples of pneumococci were analyzed whether ECVES is able to identify serotype correctly. After confirming the performance, serotyping assay was applied to identify serotypes of 528 clinical isolates of pneumococci collected from 3 different hospitals. Results: After establishment of the multibead pneumococcal serotyping assay system at ECVES, 75 test samples were analyzed. There was no discrepancy of serotypes of 75 test samples between the results assigned at UAB and those at ECVES. The serotypes of 528 pneumococci isolated from patients or healthy subjects were determined in 94.3% of isolates (498/528). Conclusions: The multibead pneumococcal serotyping assay can be successfully established in Korea. With this method, surveillance of serotypes of pneumococci isolated from patients as well as healthy subjects could be studied.

Receptor Binding Affinities of Synthetic Cannabinoids Determined by Non-Isotopic Receptor Binding Assay

  • Cha, Hye Jin;Song, Yun Jeong;Lee, Da Eun;Kim, Young-Hoon;Shin, Jisoon;Jang, Choon-Gon;Suh, Soo Kyung;Kim, Sung Jin;Yun, Jaesuk
    • Toxicological Research
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    • v.35 no.1
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    • pp.37-44
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    • 2019
  • A major predictor of the efficacy of natural or synthetic cannabinoids is their binding affinity to the cannabinoid type I receptor ($CB_1$) in the central nervous system, as the main psychological effects of cannabinoids are achieved via binding to this receptor. Conventionally, receptor binding assays have been performed using isotopes, which are inconvenient owing to the effects of radioactivity. In the present study, the binding affinities of five cannabinoids for purified $CB_1$ were measured using a surface plasmon resonance (SPR) technique as a putative non-isotopic receptor binding assay. Results were compared with those of a radio-isotope-labeled receptor binding assay. The representative natural cannabinoid ${\Delta}^9$-tetrahydrocannabinol and four synthetic cannabinoids, JWH-015, JWH-210, RCS-4, and JWH-250, were assessed using both the SPR biosensor assay and the conventional isotopic receptor binding assay. The binding affinities of the test substances to $CB_1$ were determined to be (from highest to lowest) $9.52{\times}10^{-3}M$ (JWH-210), $6.54{\times}10^{-12}M$ (JWH-250), $1.56{\times}10^{-11}M$ (${\Delta}^9$-tetrahydrocannabinol), $2.75{\times}10^{-11}M$ (RCS-4), and $6.80{\times}10^{-11}M$ (JWH-015) using the non-isotopic method. Using the conventional isotopic receptor binding assay, the same order of affinities was observed. In conclusion, our results support the use of kinetic analysis via SPR in place of the isotopic receptor binding assay. To replace the receptor binding affinity assay with SPR techniques in routine assays, further studies for method validation will be needed in the future.

Comparison of Isolation Agar Method, Real-Time PCR and Loop-Mediated Isothermal Amplification-Bioluminescence for the Detection of Salmonella Typhimurium in Mousse Cake and Tiramisu (Mousse cake와 Tiramisu에 인위접종된 Salmonella Typhimurium의 식품공전 분리배지, Real-time PCR과 Loop-mediated isothermal amplification-bioluminescence의 검출 특성 비교)

  • Lee, So-Young;Gwak, Seung-Hae;Kim, Jin-Hee;Oh, Se-Wook
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.290-295
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    • 2019
  • Salmonella spp. are frequently associated with food and are among the most important foodborne pathogens. The recent Salmonella out breaks in Korea was associated with chocolate mousse cakes served with school meals during September 2018. The objective of this research was to compare the 3M Molecular Detection Assay 2 - Salmonella and the Korean Standard Method of Salmonella in artificially inoculated mousse (chocolate and cheese) and tiramisu cakes. Mousse (chocolate and cheese) and tiramisu cakes were artificially inoculated with S. Typhimurium. Twenty five gram of sample was enriched with 225 mL buffered peptone water for incubation at $37^{\circ}C$ for 24 h. After enrichment, the cultures were analyzed by using the 3M Molecular Detection Assay 2 - Salmonella and the Korean Standard Method. Most of the inoculated samples showed similar results except the chocolate mousse cakes, in which real-time PCR was unable to detect S. Typhimurium even after $10^4CFU/25g$ of inoculation. However, S. Typhimurium inoculated at a concentration of $10^0CFU/25g$ was detected by using 3M Molecular Detection Assay 2 - Salmonella. In chocolate mousse, detection of S. Typhimurium using real-time PCR was partially successful when dark chocolate was added at less than 15%. Negative results in real-time PCR and 3M Molecular Detection Assay 2 - Salmonella were confirmed by gel electrophoresis. The data indicated that dark chocolate could inhibit amplification of the target gene in the PCR reactions. In conclusion, the 3M Molecular Detection Assay 2 - Salmonella was better than the Korean Standard Method (real-time PCR) for the detection of S. Typhimurium in chocolate mousse cakes and chocolate mousse.

Evauation of Hygienic Status of High School Foodservice using ATP Bioluminescence Assay & Microorganism Test Strips (ATP Bioluminescence와 간이미생물배지를 이용한 고등학교 급식시설의 위생상태 조사연구)

  • Jang, Jae-Seon;Hwang, Seong-Hee
    • The Korean Journal of Food And Nutrition
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    • v.28 no.5
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    • pp.918-925
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    • 2015
  • An investigation was conducted to evaluate the hygienic status of 33 high school foodservice systems in Yongin city by using hygiene management guide checklist, ATP bioluminescence assay and microbe inspection petrifilm (APC, coliform group, Staphylococcus aureus) of food utensils during use. The 22 hygiene management guide checklist items about facilities, personal hygiene, food control, distribution, washing and disinfection had good grade but there were some inadequate behaviors on observation. The inspection results showed their sanitary condition met the level B of the recommendation of Korea method, it means sanitary management system get settled but more practical CCP system was needed. ATP bioluminescence assay was conducted on surface of food facilities, ATP ranged 425~2,552 RLU on gloves, 541~70,251 RLU on apron, 1,596~88,490 RLU on working desk, 1,177~263,813 RLU on sterilizer grip, 715~32,814 RLU on sterilizer shelf, 114~619,725 RLU on refrigerator grip, 677~319,007 RLU on refrigerator shelf, 71~196,725 RLU on freezer grip, 1,535~233,375 RLU on freezer shelf. APC ranged $66.7{\pm}29.0CFU$ on freezer grip, $102.1{\pm}35.9CFU$ on refrigerator grip, $45.4{\pm}28.2CFU$ on heating cabinet grip, $58.8{\pm}40.4CFU$ on sterilizer grip, the number of coliform group ranged $5.6{\pm}4.9CFU$ on freezer grip, $9.1{\pm}8.7CFU$ on refrigerator grip, $1.2{\pm}1.1CFU$ on heating cabinet grip, $4.5{\pm}4.4CFU$ on sterilizer grip. S. aureus ranged $8.0{\pm}5.6CFU$ on freezer grip, $12.2{\pm}9.6CFU$ on refrigerator grip, $2.1{\pm}1.6CFU$ on heating cabinet grip, $11.6{\pm}6.4CFU$ on sterilizer grip.

Development of Heterologous ELISA System for Diagnosis of Pregnancy in Swine (돼지 임신 진단을 위한 Heterologous ELISA 법 개발)

  • Park, Sung-Min;Lee, An-Na;Yoon, Taek-Joon;Park, Yong-Su;Song, Tae-Jun;Kim, Young-Hoon;An, Hyo-Sun;Park, Won-Choul;Kim, Jong-Bae
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.161-167
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    • 2010
  • Early diagnoses of pregnancy for animal such as swine and bovine is extremely important to increase income of a farmhouse and for the management of farm. For the development of immunoasaay system of pregnancy in swine, we report a competitive heterologous enzyme linked immunosorbent assay (ELISA) for the direct measurement of oestrone sulfate (E1S) in diluted urine using anti-E1G (glucuronide) monoclonal antibody which cross react with ElS. The principle of assay was based on the typical solid-phase competitive ELISA methods using E1G-HRP (horseradish peroxidase) as a tracer and E1S for standard. The method had a reasonable sensitivity for the detection of E1S with 0.15 ng/ml as a detection limit. The intra-assay and inter-assay precisions were raging coefficient of from 8.50~9.67% and 8.50~9.87%, respectively, which were quite acceptable. In a field trial with a group 37 sows (18 non-pregnancy and 19 pregnancy sows) after day 29~30 post service, the concentration of E1S were determined to be below 30 ng/ml in all non-pregnancy group and over 48 ng/ml in pregnancy group except one sample. The method described here, heterologous ELISA for the measurement of E1S in urine is good enough for monitoring the early pregnancy test of swine.